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Woodford, R.

Publications and source records attributed to Woodford, R..

8 recordsLinked to original sources

PsbS confers limited adaptive benefit to C4 photosynthesis under fluctuating light

Adaptation of plant photosynthesis to dynamic light conditions experienced in natural environments is achieved through specific protective mechanisms. Energy-dependent non-photochemical quenching (qE), regulated by Photosystem II Subunit S (PsbS), is a key process facilitating acclimation to fluctuating light in C3 plants, which operate conventional photosynthesis. C4 plants, which include some of the worlds most productive and agriculturally important crops, have evolved a distinct high-efficiency photosynthetic pathway. Little is known about the role of specific processes, like qE, in acclimation of C4 plants to dynamic light environments. We generated gene-edited lines of the model C4 grass Setaria viridis lacking PsbS, which were found to be deficient in qE. This deficiency resulted in a modest increase in PSII photoinhibition and a CO2 assimilation penalty under light stress in short-term experiments, but photosynthesis and growth under fluctuating light were unaffected. Instead, keeping Photosystem I oxidised through photosynthetic control, negative feedback regulation of the Cytochrome b6f complex, was critical. Therefore, unlike in C3 plants, qE does not provide a significant adaptive advantage to C4 plants under dynamic light conditions. These findings provide important insights into the biology of C4 plants and help prioritise future strategies for improving the productivity and resilience of C4 crops.

plant biology↗

The Role of Phosphoenolpyruvate Carboxylase-Protein Kinase in C4 Photosynthesis: Insights from Zea mays Mutant Analysis

Understanding the regulation of enzyme activity involved in photosynthesis is essential for engineering enhanced carbon fixation in crops. In C4 plants, the enzyme phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is one of the most abundant leaf enzymes and plays an essential role in photosynthetic carbon dioxide (CO2) fixation. The enzyme also plays a key role in central metabolism (e.g., providing intermediates to the citric acid cycle) and therefore must be highly regulated to coordinate its activity. The regulation of PEPC activity can occur allosterically by glucose 6-phosphate activation and malate inhibition, which is in part influenced by reversible phosphorylation. A specific light-dependent phosphorylation of PEPC at an N-terminal serine residue by the PEPC-protein kinase (PEPC-PK) can regulate its sensitivity to this allosteric regulation. However, the impact of this PEPC phosphorylation has not been tested in a C4 crop. Therefore, we created PEPC-PK mutant lines in Zea mays to assess the impact of PEPC phosphorylation on its allosteric regulation, photosynthesis, and growth. While the maximum PEPC activity was unchanged, PEPC in the PEPC-PK mutant plants was not phosphorylated under light and was more sensitive to malate inhibition. However, gas exchange, electron transport, and field biomass analyses showed no differences in the PEPC-PK mutant plants. These results demonstrate that in Z. mays PEPC phosphorylation affects enzyme sensitivity to malate in vitro but does not substantially alert photosynthetic performance or growth under field conditions suggesting additional regulation of PEPC activity in planta.

plant biology↗

Fitting photosynthetic carbon dioxide and irradiance response curves for C4 leaves

Gas exchange measurements provide crucial insights into the complex mechanisms of photosynthesis. Responses of CO2 assimilation rate (A) to intercellular CO2 partial pressure (Ci) and irradiance (I) link gas exchange measurements to the underlying photosynthetic biochemistry of a leaf. The unique biochemistry and leaf anatomy which distinguish C4 photosynthesis make it necessary to apply models and fitting routines which appropriately parameterise and incorporate these characteristics. Here we provide updates to the C4 photosynthesis model by improving the parameterisation of cyclic electron flow in C4 photosynthesis using experimentally derived values from S. viridis. We additionally describe two fitting routines for assessing C4 photosynthesis based on the updated model. Fitting of a CO2 response curve (A/Ci) provides estimates of maximum PEP carboxylase activity (Vpmax) and maximum Rubisco activity (Vcmax), and calculates the electron transport rate (J) needed to sustain the measured CO2 assimilation rate (A). Fitting of an irradiance response curve (A/I) provides estimates for the maximum electron transport rate (Jmax), day respiration rate (Rd), the quantum yield ({phi}CO2) and light compensation point ({Gamma}light). Values of the above output parameters are provided at both the measurement temperature and at 25 {degrees}C for ease of comparative reporting. The fitting tool has been designed in Microsoft Excel to minimise barriers to entry and enable simplicity of fitting while simultaneously catering to individuals with diverse expertise and experience in C4 gas exchange modelling.

plant biology↗

PGR5 is needed for redox-dependent regulation of ATP synthase both in chloroplasts and in cyanobacteria

The regulation of proton motive force (pmf) via ATP synthase activity is a critical mechanism by which photosynthetic organisms maintain redox homeostasis and activate photoprotective responses under fluctuating light conditions. Here, we used time-resolved electrochromic shift (ECS) measurements to investigate pmf dynamics across a range of photosynthetic organisms. Our results reveal that ATP synthase is dynamically regulated during light fluctuations, but in Arabidopsis thaliana could this regulation could not be explained by the known light-induced reduction of the CF{square}{gamma} subunit, suggesting alternative control mechanisms. The PROTON GRADIENT REGULATION 5 (PGR5) protein, previously proposed to facilitate cyclic electron transport (CET), also has a potential role in ATP synthase regulation. The physiological role of cyanobacterial Pgr5 has remained elusive. We characterised a {Delta}pgr5 mutant of Synechocystis sp. PCC 6803 and investigated pmf dynamics in pgr5 mutants of Chlamydomonas reinhardtii, Arabidopsis, and the C4 grass Setaria viridis. While PGR5 was not required for CET in Synechocystis, it was needed for downregulating ATP synthase under high irradiance in all tested organisms via a thiol redox state dependent mechanism. Furthermore, interaction between AtPGR5 and AtCF{square}{gamma} suggests that PGR5 functions as a conserved inhibitor of ATP synthase under high light, contributing to pmf retention and photoprotection. HighlightPGR5 is involved in regulation of ATP synthase in fluctuating light conditions across photosynthetic organisms in a manner that depends on the thiol redox state of the chloroplast or cell.

plant biology↗

PGR5 promotes energy-dependent non-photochemical quenching to enable efficient C4 photosynthesis under fluctuating light

PROTON GRADIENT REGULATION 5 (PGR5) is essential for generating proton motive force across thylakoid membranes in C3 plants and supporting photoprotection under fluctuating light conditions. It is proposed that this function is achieved by regulating cyclic electron flow around Photosystem I. During the evolutionary transition from C3 to C4 photosynthesis, the leaf abundance of PGR5 has increased, coinciding with a rise in the cyclic electron flow rate. To investigate the contribution of PGR5 to photoprotection in C4 photosynthesis, we generated model C4 monocot Setaria viridis with null pgr5 alleles. We show that plants lacking PGR5 struggle to establish proton motive force and energy-dependent non-photochemical quenching (qE) at higher irradiances during instantaneous measurements. This leads to a progressive decline in maximum Photosystem I activity when leaves are exposed to repeated cycles of high irradiance. Additionally, plants without PGR5 exhibit severely reduced growth and photosynthesis compared to wild type plants when grown under fluctuating daylight but not under constant daylight. In the absence of PGR5, a slower-relaxing, zeaxanthin-dependent form of non-photochemical quenching supports growth under fluctuating light, albeit at the cost of reduced photochemical efficiency and assimilation rate. Our findings underscore the role of PGR5 in enabling efficient C4 photosynthesis under fluctuating light by establishing proton motive force for the rapid upregulation of qE and preventing photodamage to the electron transport machinery. This study highlights the importance of various non-photochemical quenching mechanisms for C4 photosynthesis and emphasises the role of PGR5 in the evolution of C4 plants.

plant biology↗

NDH complex-mediated cyclic electron flow in bundle sheath cells enables C4 photosynthesis

The superior productivity of C4 plants is achieved via a metabolic C4 cycle which acts as a CO2 pump across mesophyll and bundle sheath (BS) cells and requires an additional input of energy in the form of ATP. Chloroplast NADH dehydrogenase-like complex (NDH) increases ATP production in C3 plants by operating cyclic electron flow (CEF) around Photosystem I (PSI), and its importance for C4 photosynthesis has been proposed from evolutionary and reverse genetics studies. We used the gene-edited C4 species Setaria viridis with null ndhO alleles lacking NDH to study a contribution of the complex to the cell-level electron transport. Our results indicate that NDH is the primary PSI electron acceptor mediating the majority of CEF in BS cells whilst the contribution of the complex to CEF in mesophyll cells is minimal. Moreover, the reduced leaf CO2 assimilation rate and growth of plants lacking the complex cannot be rescued by supplying additional CO2, indicating that NDH is essential for generating ATP required for CO2 fixation by the C3 cycle. Hereby we resolve a cell-level mechanism for the contribution of NDH to supporting high CO2 assimilation rates in C4 photosynthesis.

plant biology↗

Faster responses of photosynthesis to light transitions increase biomass and grain yield in transgenic Sorghum bicolor overexpressing Rieske FeS

Sorghum is one of the most important crops providing food and feed in many of the worlds harsher environments. Sorghum utilises the C4 pathway of photosynthesis in which a biochemical carbon concentrating mechanism results in high CO2 assimilation rates. Overexpressing the Rieske subunit of the Cytochrome b6f complex was previously shown to increase the rate of photosynthetic electron transport and stimulate CO2 assimilation in the model C4 plant Setaria viridis. To test whether productivity of C4 crops could be improved by Rieske overexpression, we created transgenic Sorghum bicolor plants with increased Rieske content. The transgenic plants showed no marked changes in abundance of other photosynthetic proteins or chlorophyll content. Increases in yield of Photosystem II and CO2 assimilation rate as well as faster responses of non-photochemical quenching during transient photosynthetic responses were observed as a result of an elevated in vivo Cytochrome b6f activity in plants overexpressing Rieske. The steady-state rates of electron transport and CO2 assimilation did not differ between transgenic and control plants, suggesting that Cytochrome b6f is not the only factor limiting electron transport in sorghum at high light and high CO2. Nevertheless, more agile responses of photosynthesis to light transitions led to increases in biomass and grain yield in plants overexpressing Rieske. Our results indicate that increasing Rieske content could boost productivity of C4 crops by improving the efficiency of light utilisation and conversion to biomass.

plant biology↗

Rieske FeS overexpression in tobacco provides increased abundance and activity of Cytochrome b6f

Photosynthesis is fundamental for plant growth and yield. The Cytochrome b6f complex catalyses a rate-limiting step in thylakoid electron transport and therefore represents an important point of regulation of photosynthesis. Here we show that overexpression of a single core subunit of Cytochrome b6f, the Rieske FeS protein, led to up to a 40% increase in the abundance of the complex in Nicotiana tabacum (tobacco) and was accompanied by an enhanced in vitro Cytochrome f activity, indicating a full functionality of the complex. Analysis of transgenic plants overexpressing Rieske FeS by the light-induced fluorescence transients technique revealed a more oxidised primary quinone acceptor of Photosystem II (QA) and plastoquinone pool and a faster electron transport from the plastoquinone pool to Photosystem I upon changes in irradiance, compared to control plants. A faster establishing of qE, the energy-dependent component of non-photochemical quenching, in transgenic plants suggested a more rapid build-up of the transmembrane proton gradient, also supporting the increased in vivo Cytochrome b6f activity. However, there was no consistent increase in steady-state rates of electron transport or CO2 assimilation in plants overexpressing Rieske FeS grown in either laboratory conditions or in field trials, suggesting that the in vivo activity of the complex was only transiently increased upon changes in irradiance. Our results show that overexpression of Rieske FeS in tobacco enhances abundance of functional Cytochrome b6f and electron transport capacity and may have a potential to increase plant productivity if combined with other traits. One-sentence summaryIncreased abundance of Cytochrome b6f complex leads to transient increases in photosynthetic electron transport rate in tobacco.

plant biology↗